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Micro/Para (dra Madrid)

Amoeba
20 January 08

Species of Amoeba • Asymptomatic cyst passer


1. Entamoeba histolytica • Non-dysenteric diarrhea
2. E. dispar 3. Amebic dysentery
3. E. hartmanni • Seldom epidemic
4. E. coli
• Gradual onset
5. E. polecki
6. Endolimax riaria • No prodromal feature, no vomiting
7. iodamoeba butschilii • Patient usually ambulant, mild abdominal cramps
• Tenesmus uncommon
Fecal-Oral Transmission Factors • Bloody diarrhea, fishy odor stool
• Poor personal hygiene • Stool microscopy: few bacilli, red cells, troph with
• Children (eg. Day care centers) ingested RBC, Charcot-leyden crystals
• Institutions (eg prisons, mental hospital, orphanages) 4. Extraintestinal disease
• Food handlers • Liver abscess – most important and most common; 1-
7% of children with massive infection; R lobe 53%,
Developing Countries both lobes 25%, L lobe 8%
• Poor sanitation • Pleuropulmonary amebiasis – 2nd most common
• Lack of indoor plumbing Secondary to direct extension from a hepatic abscess
• Endemic perforates thru the diaphragm and into a bronchus appears
• Travelers’ diarrhea as pneumonic consolidation of the lower R lung
• Brain (meningoencephalitis) – 1-2% and other organs
Water-borne epidemics • Cutaneous and genital diseases
Male homosexuality 5. Invasive
Oral-anal contact • Necrosis of mucosa – ulcer
o Dysentery
Zoonosis o Hematophogous trophozoites
• Entamoeba = no • Ulcer enlargement – peritonitis
• Cryptosporidium = yes o Occasional ameba
• Giardia = controversial • Metastasis – extraintestinal amebiasis
o Via blood stream or direct extensions
Entamoeba histolytica o Primarily liver-amebic abscess
• Pseudopod-forming nonflagellated protozoan parasite o Other sites infrequent
• Most invasive o Ameba-free stools common
• Only member of the Entamoeba family to cause colitis and
liver abscess E. histolytica forms
1. Cyst
Disease: Amoebiasis • Resistant to gastric acidity and dessication
Site: Large intestine • Can survive in a moist environment for several weeks
Portal of Entry: Mouth • Acquired by ingestion via the fecal/oral route
Modes of Transmission: Ingestion from fecally contaminated • Contains chitin
material
Others: • Resistant to chlorine and cold water
• Venereal: fecal-oral contact • Killed by heat and dessication, removed by filtration
• Direct colonic inoculation through contaminated ename • 3.5 – 15 µm
equipment • Diagnostic form: mature cyst
o Shape usually spherical
Clinical Syndromes Associated with Amoebiasis o 4 nuclei
1. Intestinal Diseases o infective stage
• Asymptomatic cyst passer • Immature cyst
• Symptomatic non-dysenteric infection o shape usually spherical
• Amebic dysentery (acute) o one large (or 2 medium sized) nucleus
• Fulminant colitis – perforation (peritonitis) most serious o Cigar/sausage shaped chromatoidal bar with
complication rounded ends
• Amoeba (amebic granuloma: resembles o Glycogen vacuoles
adenoCA); .1% sx of intestinal obtrusion 2. Trophozoites
• Active, motile feeding sage
2. Non-Invasive • Causes pathology in color
• Ameba colony on mucosa surface • Anaerobic

Isay…^_^ 1 of 5
Micro/Para – Amoeba by Dra Madrid Page 2 of 5

• No mitochondria Treatment
• Ingest RBC (pathognomonic) 2 Objective:
• Lyses tissue – necrosis 1. To cure invasive disease at both intestinal and
extraintestinal sites
• Can cause invasive disease
2. To eliminate the passage of cysts from intestinal lumen
• Cannot survive in the environment
• 10-20 u • Treatment for asymptomatic: Iodoquinol, Paramomycin, or
• clean delicate nucleus with evenly distributed peripheral Diloxanide furoate
chromatic • Treatment for nondysenteric: Dysenteric or extraintestinal-
• small central karyosome metronidazole or Inidazole + luminal agent
• “clean” cytoplasm
• ingested RBC may be present in cytoplasm Prevention and Control
• Integrated and community based efforts thru health
Possible Virulence Factors education and promotion
• Host factors (eg immune response) • Improve environmental sanitation
• Parasite factors • Provide for:
• Resistance to host response (complement resistance) o Sanitary disposal of human feces
• Adherence properties (Eh-lectin) o Safe drinking water – boiled or filtered
• Cytolytic properties (adherence + amebapore) o Safe food – thorough washing of raw fruits and veggies
• Ability of breakdown tissues (secreted protease) o Monitor food handlers
• Vaccines (animal models) – mucosal immune response
Entamoeba Prevalence recombinant amebic antigen more advantageous than
• E. dispar – 10 fold> E. histolytica inactivated/attenuated ameba: serine rich E. histolytica
protein (SREHR) adherence lectins (Gal/GalNac lectin) 29
• Discrete endemic pockets of E. histolytica observed kDa cysteine rich amebic antigen
• 25% seropositive for E. histolytica in endemic areas
COMMENSAL AMOEBA
• 10% infected with E. histolytica will develop invasive
Entamoeba Endolimax Iodamoeba
amebiasis NUCLEUS Spherical Vesicular
NUCLEAR Direct lines Anchored
Diagnosis MEMBRANE with by
DFS (fresh stool) chromatin achromatic
• With saline solution: trophozoite motility (unidirectional granules fibrils
movement) KARYOSOME Small Large, Large
center of irregular, chromatin
• Saline and methylene blue: will stain blue vs. WBC nucleus anchored rich
• Saline and iodine: observe nucleus and karyosome to nucleus surrounded
More sensitive test: by by a layer
Concentration Methods achromatic of
• Formaline Ether Concentration Test (FEGT) globules achromatic
fibrils
• Merthiolate Iodine Formalin Concentration Test (MIFO)
• Stool culture: Robinson’s and Inoki medium
Entamoeba Dispar
Intestinal infections • Morphologically similar to E. histolytica
Note: • Different DNA and ribosomal RNA
• For size of cyst • Different isoenzyme pattern
• Number of nuclei
• Location and appearance of karyosome Entamoeba Hartmanni
• Characteristic appearance of chromatoid bodies • Similar to E. histolytica except much smaller
• (+) of cytoplasmic structures (glycogen vacuoles) • Does not ingest RBC
Sigmoidoscopy – lesion, aspirate, biopsy • More sluggish in movement
Antigen detection – histolytica/dispar • Cyst Mature: 5-10 um, 4 nuclei, coarse cytoplasm
• Immature: chromatoidal bars: short with tapered ends thin
For extraintestinal (hepatic) bar like
• Serology – key in diagnosis of ALA, IHAT (also past
infection), IFAT, (CIA, AGD, ELISA – short duration, few Entamoeba Coli
months) only E. histolytica • Harmless inhabitant of the intestine
• Imaging – CT, MRI, ultrasound – 80% of cases: R lobe • Cyst
(round or oval hypoechoic area with wall echoes) o 10-30 u
• Abscess aspiration o Mature form has 8 nuclei
o Only select cases o Granular cytoplasm
o Reddish brown liquid o Chromatoid bodies when present have splintered ends
o Trophozoites at abscess wall • Trophozoites
o 20-25 um
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o Broad blunt pseudopodia • Large glycogen vacuole


o Single nucleus with eccentric chromosome Trophozoites
o “dirty” cytoplasm • 6-20 um (9-14 um long)
o Ingestion of bacteria but NOT red cells • Large vesicular nucleus with large endosome surrounded
o Sluggish, indeterminate movements by achromatic granules
• No peripheral chromatin granules on the nuclear membrane
E. coli E. histolytica • Sluggish movement
CYSTS
• Rarely seen
SIZE Larger
NUCLEI 8 4
CYTOPLASM More granular Clear Endolimax Nana
CHROMATOIDAL Splinter-like Rounded Disease: Non pathogenic
BODIES ends Site: Intestine
TROPHOZOITES Portal of Entry: Mouth
ENDOPLASM More vacuolated or Source of Infection: Ingestion of Cyst
granular Forms: Cyst
ECTOPLASM Narrower, less • 6-12 um
differentiated
PSEUDOPODIA Broader and blunter • Mature cyst has 4 nuclei
MOVEMENT Sluggish, unidirected Unidirect • Oval shape
ional • Refractile karysosomes
CHROMATIN Thicker, irregular, Evenly Trophozoites
peripheral distributed
• Approx. 15 u at most
KARYOSOME Large, eccentric Small
• Nucleus: large, irregular karyosome
• Thick nuclear membrane
Entamoeba Polecki
• Without chromatin beading
• Parasite of pigs and monkeys
• Sluggish motility
• Rarely may infect humans
• Blunt hyaline pseudopods
• Cyst consistently uninucleated
• Stained smear: prominent nuclear membrane and
Blastocystis Hominis
karyosome
• Inhabitiant of the lower intestinal tract of humans and other
animals
Entamoeba Gingivals
• Formerly classified as yeasts
• Light microscopy has since shown that it lacks a
• Lives on gums and teeth surface, gum pockets, tonsillar mitochondrion with a protozoan morphology
crypts • Capable of pseudopodial extension and retraction
• Exhibits similar morphology to E. histolytica
• Reproduction is asexual through binary fission or
• Transmission: sporulation under strict anaerobic conditions
o Direct by kissing, droplet spray, sharing utensils
o Can multiply in bronchial mucus Parasite Biology
o No cysts stage • Multiplication is by binary fission
o Trophozoites: ingested leukocytes • Transmission: Fecal oral route
 10-20 um • Morphological forms:
 Moves quickly, numerous blunt pseudopodia o Vacuolated
 Food vacuoles: numerous cellular debris and o Ameba-like
bacteria o Granular
 Recovered from patients with periodontal diseases o Multiple fission
 Non pathogenic o Cyst
o Avacuolar forms
Diagnosis: Swab between the gums and teeth (+) for
trophozoites
Vaculoated forms
Treatment: None
• Most predominant forms in the stool
Iodamoeba Buschlii • Spherical, measures 5-10 um in diameter
Disease: Non Pathogenic • Large central vacuole (reproductive organelle) pushes the
Site: Intestine cytoplasm and nuclei on the periphery of the cell
Portal Of Entry: Mouth • Sometimes a thick capsule surrounds the vacuolated form
Source of Infection: Ingestion of Cyst • Are the main type of Blastocystis causing diarrhea
Forms: Cyst
• 10-12 um (5-20) Ameba-like forms
• Shape: ovoidal or rounded • Occasionally observed in the stool
• Uninucleated • Exhibit active extension and retraction of pseudopodia
• Large eccentric karyosome • Nuclear chromatin shows peripheral clumping
• (-) chromatoid bodies
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• An intermediate stage between the vacuolar form and the • Hides in intestinal mucus, sticks and holds on to intestinal
cystic form membranes
• DOC: metronidazole, iodoquinol, TMP-SMX
Granular forms
• Mainly observed from old cutlrues Epidemiology
• Diameter: 10-60 um • Worlwide: common in tropical, subtropical, and developing
• Granular contents develop into daughter cells of the countries
ameba-form when the cell ruptures • All ages are affected but asymptomatic cases are
• Multiple fission: arise from vacuolated forms commonly found in children and in host with weakened
immune system
Cystic form • Infection more likely to occur in crowded and unsanitary
• Size: 3-55 um conditions
• Very prominent and thick osmophilic electron dense wall • Philippine prevalence of 20.7% in 772 stool samples
• Appears as a sharply demarcated polymorphic, oval or examined in 1998
circular, dense body surrounded by a loose outer • Studies show prevalence rates of 40.6% among food
membranous layer service workers in tertiary hospitals and 23.6% among food
• It is assumed that the thick walled cyst might be handlers in selected school canteens in Manila
responsible for external transmission while those cysts with • Stool surveys done by FETP of the DOH
thin walls might be the cause of reinfection within a host’s o Tapel Gonzage, Cagayan Valley: 20% prevalence rates
intestinal tract o Talavera, Nueva Ecija 44%
• Several animals (macaques, chicken, dogs, ostriches)
Clinical Manifestations harbor the parasite; also present in house lizards and
• Several studies showed that presence of B. hominis was cockroaches food and water contaminated by their fecal
not associated with symptoms or is found with other droppings may transmit Blastocytis
organisms that are more likely to be the cause of symptoms
• Other studies concluded that in large numbers, B. hominis Prevention and Control
produce a wide variety of intestinal disorders • Safe drinking water
• Among immunocompromised subjects, B. hominis showed • Sanitary preparation of food
a significant association with GI symptoms • Cysts of Blastocystis can survive up to 49 days in water at
• Symptoms usually last for 8-10 days but may persist for normal temperature and have shown resistance to chlorine
weeks or months at the standard concentrations
o Abdominal cramps
o Irritable bowel syndrome
o Bloating Thanks isay sa pagtype nito for me…love you! Mwah..!!! hehe…
o Flatulence happy aral sa inyo…^_^
o Mild to moderate diarrhea without fecal leukocytes or
blood ----Shar
o Vomiting
o Low grade fever
o Malaise

Diagnosis
• Stool samples collected more than once
• Direct fecal smear – sensitivity increased when
concentration techniques are used
• Hematodin or trichome staining – differentiate the various
stages of Blastocystis
• Leukocytes are usually seen in fecal smear and stool
eosinophilia may be observed
• Organism can be cultured using the Boeck and Orbohierv’s
or the Nelson and Jones media
• Permanently stained smears are preferred over the mount
preparations because fecal debris maybe mistaken for the
organisms
• Do not wash specimens in water, as this will lyse the
organisms resulting in false negatives
• Microscopy: B. hominis appears as spherical or oval cyst-
like structures

Treatment
• Difficult to eradicate
Micro/Para – Amoeba by Dra Madrid Page 5 of 5

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